The mission of the National Resource for Aplysia at the University of Miami (the Resource) is to produce and distribute consistently high quality cultured sea hares, Aplysia californica, for biomedical research and education and to improve the usefulness of this model system by applied research. The two aims of the current phase (years 24 ? 28) of this resource program are: 1. Resource core - Production and Distribution ? The Resource will continue to culture A. californica and make all life stages, from eggs to large adults, available to researchers at a reasonable cost. We anticipate sales will continue at a level of 10,000 ? 15,000 animals per year. We will also accommodate requests by users for separately reared, same-aged cohorts, specialized diets, or harvesting of specific tissues for RNA or DNA. In addition, we will make available mature and senescent stage animals, validated using criteria generated by the Resource, as well as additional specialized animals as developed in our planned Applied Research programs. 2. Applied Research Programs to support the Resource ? We will conduct two applied research projects to create new Aplysia resources that we believe will expand the usefulness of this model system to researchers: 2.1 Molecular characterization of the caloric restriction model of arrested aging ? We will investigate the mechanisms involved in lengthening the one year Aplysia lifecycle via caloric restriction (CR). We will document the portions of the life cycle such as pre-reproductive, mature and aged that are prolonged by CR. We will also examine alterations in previously identified gene expression profiles associated with aging at these stages, and determine how they are affected by CR. This project will expand the usefulness of these animals as models of aging. 2.2 Molecular characterization of the hypoxia-resistant nervous system of Aplysia ? Hypoxia and ischemia tolerant species have been very useful for understanding the mechanisms by which such animals avoid the catastrophic nervous system damage encountered by humans subject to stroke or similar injuries. Aplysia are very resistant to hypoxia such that their simple, well studied nervous system should be an ideal model for investigating transcriptomic and metabolic adaptations that make this resistance possible. This would create a novel model for study of mitigation of hypoxic damage in the nervous system. The requested administrative supplement will provide funds for the purchase of critical equipment to support these ongoing programs. Specifically: A qPCR machine is urgently needed to support projects underway in the Resource core (detection of viral loads in tissues of Aplysia, both those reared at the hatchery as well as those from the wild) and Aim 2.2 of the applied research program (measurement of expression of specific genes associated with the hypoxia response in Aplysia). For the past five years we have been using a departmental qPCR machine which is now over-subscribed and out of date (no longer eligible for a service contract). Thus, we no longer have reliable access to this technology and urgently need to acquire a qPCR instrument dedicated to the resource. Three key pieces of equipment are needed to restore our two integrated systems for electrophysiology back to a functioning state. These integrated systems are critical to measuring neuronal cell function as part of Aim 2.1 in the applied research program. Both rigs were assembled in the 1990s and the equipment in question, one patch clamp amplifier and two sets of micromanipulators are no longer serviced by the manufacturer. Thus we urgently need funds to replace these broken instruments. We are also requesting funds for two key microscopy components in the Resource. A modern stereo microscope is needed to replace the 25 year-old model used to evaluate and measure Aplysia veliger larvae throughout their development and a low light, sCMOS, microscope camera is requested for scoring a variety of fluorescent markers in cells.